title: New methods for inducing the differentiation of amniotic-derived mesenchymal stem cells into motor neuron precursor cells.
abstract: Our study investigates the differentiation of amniotic-derived mesenchymal stem cells (ADMSCs) into motor neuron (MN) precursor cells induced by a combination of extracellular matrix (ECM) and multi-cell factors. Membrane-like ECM was made by an enzymatic and chemical extraction method and exhibited good biological compatibility. Cells in the experimental group (EG) were treated with ECM and multi-cell factors in a multi-step induction process, while the control group (CG) was treated similarly, except without ECM. In the EG, after induction, the cells formed processes that connected with neighboring cells to form a net that had directionality. In these cells, neuron-specific enolase (NSE) and synaptophysin (SYN) expression levels increased and glial fibrillary acidic protein (GFAP) expression decreased. The SYN expression in the EG cells was higher compared with those in the CG. In the CG, NSE expression increased, while the expression of Nestin and SYN did not change. These were several changes in the levels of other genes: ADMSCs at passage 1 expressed Nanog, SOX2, octamer-binding transcription factor 4 (OCT4) and Nestin. In the EG, at the beginning of induction, the expression of Nanog decreased and that of SOX2 and Nestin increased. After 2 days, the cells expressed Nestin, OCT4 and SYNIII, and after 3 days, they expressed Olig2, OCT4, Nestin, SYNII and Islet1 (ISL1). Finally, at day 6, the cells expressed Nestin, SYNI, SYNIII, ISL-1, homeobox 9 (Hb9) and oligodendrocyte lineage transcription factor 2 (Olig2). In the CG, the cells never expressed SYNI, SYNII or Hb9. Our studies therefore demonstrate that the extracted ECM was capable of promoting the maturation of synapses. Human ADMSCs are composed of multiple cell subsets, including neural progenitor cells. The multi-step induction method used in this study causes human ADMSCs to differentiate into MN precursor cells.
authors.affiliation: Department of Neurosurgery, The First People's Hospital of Jiujiang, Jiujiang, Jiangxi Province, People's Republic of China. zzdxhuw@163.com
authors.forename: Wei
authors.initials: W
authors.lastname: Hu
authors.forename: Fang-xia
authors.initials: FX
authors.lastname: Guan
authors.forename: Yuan
authors.initials: Y
authors.lastname: Li
authors.forename: You-jia
authors.initials: YJ
authors.lastname: Tang
authors.forename: Feng
authors.initials: F
authors.lastname: Yang
authors.forename: Bo
authors.initials: B
authors.lastname: Yang
mesh_terms: D000650:Amnion / Q000166:cytology; D002454:Cell Differentiation / Q000502:physiology*; D019070:Cell Lineage; D002478:Cells, Cultured; D005109:Extracellular Matrix / Q000502:physiology; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000502:physiology*; D009046:Motor Neurons / Q000166:cytology* / Q000378:metabolism; D064231:Nestin / Q000096:biosynthesis; D010751:Phosphopyruvate Hydratase / Q000096:biosynthesis; D013234:Stem Cells / Q000166:cytology*
chemical_list: D064231:Nestin; D010751:Phosphopyruvate Hydratase
keywords: AChE; ADMSCs; ASCs; Amniotic-derived mesenchymal stem cells; BDNF; CG; ChAT; DMSO; Differentiation; EG; ESCs; Extracellular matrix; FCS; GFAP; H&E; Hb9; MAPCs; MN; MN precursor cells; NF; NSE; NTs; OCT4; Olig2; PBS; RA; SYN; acetylcholine esterase; adult stem cells; amnion mesenchymal stem cells; bFGF; basic fibroblast growth factor; brain derived neurophic factor; choline acetyltransferase; control group; dimethyl sulfoxide; embryonic stem cells; experimental group; fetal calf serum; glial fibrillary acidic protein; hematoxylin and eosin; homeobox 9; motor neuron; multi-potent adult progenitor cells; neurofilament protein; neuronspecific enolase; neurotrophic factor; octamer-binding transcription factor4; oligodendrocyte lineage transcription factor 2; phosphate buffer saline; retinoic acid; synaptophysin-1